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Differential-Display Reverse Transcription-PCR (DDRT-PCR)

  • Book
  • © 1998

Overview

Authors:
  1. Sergio Colonna-Romano

    1. International Institute of Genetics and Biophysics, National Research Council, Naples, Italy

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  2. Antonella Leone

    1. Research-Institute of Vegetable and Ornamental Plant Breeding, National Research Council, Portici, Italy

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    You can also search for this author in PubMed   Google Scholar

  3. Bruno Maresca

    1. International Institute of Genetics and Biophysics, National Research Council, Naples, Italy

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    You can also search for this author in PubMed   Google Scholar

  • First manual describing this new technique
  • Easy to follow step-by-step instructions
  • Practical hints and experimental strategies are given for every protocol

Part of the book series: Springer Lab Manuals (SLM)

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Table of contents (11 chapters)

  1. Front Matter

    Pages I-X
    Download chapter PDF

  2. Overview

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 1-14

  3. Preparation of Total RNA

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 15-33

  4. Differential Display

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 34-45

  5. Size Separation of cDNA Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 46-54

  6. Isolation of Differentially Expressed cDNA Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 55-58

  7. Reamplification of Eluted cDNAs

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 59-63

  8. Cloning of Amplified cDNA Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 64-76

  9. Checking Subcloned Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 77-80

  10. Confirmation of Differential Expression of Cloned cDNA Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 81-95

  11. Northern Blot Affinity Capturing of cDNA

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 96-101

  12. Sequencing of Differentially Expressed cDNA Fragments

    • Sergio Colonna-Romano, Antonella Leone, Bruno Maresca
    Pages 102-109

  13. Back Matter

    Pages 110-124
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Keywords

About this book

Identification of differentially expressed genes is one of the major challenges in molecular biology. Several techniques allow the cloning of such sequences. However, methods such as RNA subtraction or differential hybridization are time-consuming and require large amounts of mRNA.
Recently, a new approach has successfully been developed: Differential-Display Reverse Transcription-PCR (DDRT-PCR). This technique has been proven to be highly effective in identifying sequences that are differentially expressed in various cell types. The most striking advantage is, however, that only nanograms of total RNA are sufficient. Thus every mRNA species expressed in the cell system can be investigated, even those present at very low levels.

Authors and Affiliations

  • International Institute of Genetics and Biophysics, National Research Council, Naples, Italy

    Sergio Colonna-Romano, Bruno Maresca

  • Research-Institute of Vegetable and Ornamental Plant Breeding, National Research Council, Portici, Italy

    Antonella Leone

Bibliographic Information

  • Book Title: Differential-Display Reverse Transcription-PCR (DDRT-PCR)

  • Authors: Sergio Colonna-Romano, Antonella Leone, Bruno Maresca

  • Series Title: Springer Lab Manuals

  • DOI: https://doi.org/10.1007/978-3-642-80454-0

  • Publisher: Springer Berlin, Heidelberg

  • eBook Packages: Springer Book Archive

  • Copyright Information: Springer-Verlag Berlin Heidelberg 1998

  • Softcover ISBN: 978-3-540-63297-9Published: 23 January 1998

  • eBook ISBN: 978-3-642-80454-0Published: 06 December 2012

  • Edition Number: 1

  • Number of Pages: X, 124

  • Number of Illustrations: 74 b/w illustrations

  • Topics: Cell Biology, Biochemistry, general

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